梅花品种资源叶绿体基因组SSR研究

<p>»29 9 &lt;12007 M1 Ø &nbsp; &lt; v Ð Ð JOURNALOFBEIJINGFORESTRYUNIVERSITYVol.29, Supp.1Jan., 2007là ° ù:2006-11-16htp: journal.bjfu.edu.cnÁ “:SE1 SÐÁ&#39;ù “(300070641)、SE &lt; S/?ZÏ&#39;ù “(2005004)。»BT: g ö,p V,¬ q。ö1ùîZ_:ó ±þ&#39;÷ÐâÕ。ÈÅ:010-82376017-601 Email:Luyingminbjfu.edu.cn ¹¶:100083Ø &lt;vÐ10|Q。¼½ ÖÕ&#39;÷= s8yFSSRùî吕英民1, 2 殷 婧1 杨 果1 张启翔1,2(1Ø &lt;vÐó Ðý 2SE½ñýñ/ ùîÏ)K1:¾ùî æ¨ ñÊ¥B 1õ¤¥= s8DNA(cpDNA)SSRþ(NTCP9) 948z ¼½ ¥yFDNA,É= s8DNASSRS: k。48ñ ¼½ ÖÕ¥cpSSR·Õ &nbsp;¹$,¨NTSYSpc2.11 Èq9ØNei&#39;s(1972)L. Ö“ 、e Ä ¥“ (SM)、Jaccard Ä ¥“ Dice Ä ¥“ ,isYÉ Ë、 ½s。V¥s) ¿cpDNAsÄ£ U¥ ¼½ ÖÕW¥“dÄ1“。1oM: ¼½,cpSSR,·Õm öL Ying-min1, 2;YINJing1;YANGGuo1;ZHANGQi-xiang1, 2.cpSSRanalysisofPrunusmumeculti-varresources.JournalofBeijingForestryUniversity(2007)29 (Supp.1)48-53Ch, 22 ref.1 ColegeofLandscapeArchitecture, BeijingForestryUniversity, 100083, P, R.China;2 ChinaNationalCenterforFlowerEngineeringTechnique, Beijing, 100083, P, R.China.Forty-eightcpDNAfingerprintsofPrunusmumecultivarswerebuiltupwiththeprimerNTCP9 fromtobaccocpDNAbyusingthecpSSRtechnique.BasedoncpSSRfingerprintsdataof48accessionsofP.mumecultivars, thecomplexityofrelationshipandoriginationofP.mumecultivarswereanalyzedbyusingNTSYSpc2.11toclusterandarangementanalysisofNei&#39;s(1972)geneticdistance, SMcoefi-cient, JaccardcoeficientandDicecoeficient.Keywords Prunusmume, cpSSR, DNAfingerprint®¿= s8DNA(chloroplastDNA, cpDNA)µÚ¥M &nbsp;o,yN ¼½(Prunusmume)cpDNAÐ ð±þ¥cpDNA µ÷。 Ù ð±þ  ¾ (Pinus)、Ö Ü(Zeamays)ϵZ 륡 1-15 , ¼½Ï,Ãnµ1cpDNASSRs¥¡。Ð5cpDNASSR¥ ÿ ÂDNASSR¥ ÿ &lt; 16 , V_© 、Õ、 ÖÕcpDNA¥µslM, uY Tø1“Í¥yZ ëµ +µ¥ª 。&#39;ùî ñÊB 1õ¤(Nicoti-anatabacum)¥cpDNASSRþ 948z ¼½ ¥yFDNA,ÉcpDNASSRS: k、。V¥s) ¿cpDNAsÄ£ U¥ ¼½ ÖÕW¥“dÄ1“,F &lt; ¼ ÖÕ&#39;÷¥ùî、 ¹ æ¨,# ¼½ÕÉ&#39;÷ ¹¥s0S:ùî Ø $,ÉBy ¼L. õ ím ö、s0S:£ùâÕ、×1y¥çÊÐ X H©ùîýTµBç¥ Ø I L=¨N´。1 材料与方法1.1 &nbsp;¼½ ÖÕ k cpSSR k“ Ö(nV1) 1 2Ø ¼½ öóq ¼½Ï 48ñ ¼½ ÖÕ。1.2 &nbsp;kÐsZE1.2.1 试验仪器及试剂&#39; k î P¨¥N (®ÏS j&lt; SÐý v½ñ î ÖÉ î L i4。þ#¤h1Ø Øpù½ 3þµK ³、Z 3ý 3þýñ/ µK ³。 = M ½EcoRI MseI、T4 õ¤ ½1 g 3þ/ ³。Taq ½1¿ Å 3þýñ ³、ØåS 3þ/ µK3 © ³。 &nbsp;ÖA &nbsp;TA:ØåS 3þ/ µK3 © ³。9 &lt;1 g ö©: ¼½ ÖÕ&#39;÷= s8yFSSRùî49 ðÄÐ k4:SÁsB。V1 &nbsp;¼½cpSSR k“ Ö¨:VTABLE1 SerialnumberofP.mumesamplesforcpSSRI| ÖÕ I| ÖÕ I| ÖÕ 1 lÖÜ&#39; 17 e£&#39; 33 Öî&lt;¥&#39;2 m öâ&#39; 18 B×e£&#39; 34 x½äõ&lt;¥&#39;3 v sl&#39; 19 ëÀ&#39; 35 äõ sl&lt;¥&#39;4 sl&#39; 20 õ*â½&#39; 36 5&lt;¥&#39;5 * sl&#39; 21 v½&#39; ¼37 Ö&lt;¥&#39;6 õK&#39; 22 B Ä&#39; 38 &lt;¥&#39;7 ϯK¥&#39; 23 õv £&#39; 39 C Ê&lt;¥&#39;8 ´&#39; 24  ®&#39; 40 ª&#39;9 A+&#39; 25 2Ø*£&#39; 41 Ö×Õ&#39;10 *x&#39; 26 FÜKl&#39; 42 %¥ Â=&#39;11 v&#39; 27 =U&#39; 43 ½ Â=&#39;12 äõ ûî&#39; 28 x&#39; 44 1&lt;¥&#39;13 2ë é&#39; 29 ¥&#39; 45 K Å ¦&#39;14 $ T&#39; 30 Ö &#39; 46 ö&lt;¥&#39;15 x&lt;¥&#39; 31 Ø ÖÜ&#39; 47 Æ 3&lt;¥&#39;16 Å&lt;¥&#39; 32 Ö×7&#39; ¼48 l£É¶&#39;1.2.2 引物筛选&#39; k¨13 ÷¿Ö Ü、õ¤、 ¾ = s8DNAcpSSRþ 3, 17-19 , 9 Û |¥5z ¼½ ¥yFDNAÉ+sþ ñÊ, ñʲTϵ8_© ÿ,61.54%,KÔʨ 9Kz¥B 1õ¤¥= s8DNASSRþ(NTCP9) ¼½yFDNAÉ= s8DNASSRS: k。õ¤NTCP9þ½ :5-CTTCCAAGCTAACGATGC-35-CTGTCCTATCCATTAGACAATG-31.2.3 试验方法及试验数据的记录分析方法¨F.JamesRohlf(2001)NTSYSpc2.11 Èq9 ØM »“ ÐL. Ö“ 。L. Ö“ dij¨Nei&#39;s(1972) Ö“ 。Ä ¥“ ¨SM“ &#39;e Ä ¥“ :SSM=(a+d) (a+b+c+d)J“ (Jaccard“ ):SJ=a (a+b+c)Dice“ :SC=2a (2a+b+c)TÏ,aV U ñOTUI ¥¹“1”¥ñ ;bV UBñOTU¹“1”, 6BñOTU¹“0”¥ñcV UBñOTU“0”, 6BñOTU“1”¥ñdV U ñOTU¥¹“0”¥ñ 。SAHNClusteringÉdF î Ø Ü (E(UPGMA) Ës,9 ØM »“ r,i¨MX-COMPMatrixcomparisonÏ¥COPHprogramñ½É2»Manteltest_,9 Ø“ 20 (&#39; ˲Tñ¥M1“ )rij。2 结果与分析2.1 &nbsp;¼½ ÖÕcpSSR·Õm ö# Ës= s8DNA¥±ÏsöV¥£ U¼½ ÖÕW¥ Tø1“。Ü ñÊ P¨¥þ 915HSSR ÿ,: c Ë Â/。m1 þNTCP9·ÕmFIGURE1 cpSSRfingerprintofprimercombinationsNTCP92.2 &nbsp;¼½ ÖÕcpDNASSRM » Ë®cpSSR+sþNTCP9¤¥·Õm ö(nm1): c²TÉ Ës,¤ m。1)¨Nei&#39;s(1972)L. Ö“ ¤¥ Ë Âm2 î U。2)¨e Ä ¥“ (SM)¤¥ ˲T Âm3 î U。3)¨Jaccard Ä ¥“ ¤¥ Ë Âm4î U。4)¨Dice Ä ¥“ ¤¥ Ë Âm5î U。5)1 4Õ Ës²T Âm6 î U。50 Ø &nbsp; &lt; v Ð Ð »29 m2 ¨Nei&#39;s(1972)L. Ö“ ¤¥ Ë FIGURE2 cpSSRdendrogramofNei&#39;s(1972)geneticdistancecoeficientsm3 ¨SM Ä ¥“ ¥ Ë FIGURE3 Dendrogramofsimplematchingcoeficients(SM)m4 ¨Jaccard Ä ¥“ ¤¥ ËmFIGURE4 DendrogramofJaccardcoeficientsm5 ¨Dice Ä ¥“ ¤¥ ËmFIGURE5 DendrogramofDicecoeficients9 &lt;1 g ö©: ¼½ ÖÕ&#39;÷= s8yFSSRùî51 m6 &nbsp;˲T1 mFIGURE6 MXCOMPmatrixcomparisonofdendrograms52 Ø &nbsp; &lt; v Ð Ð »29 2.3 &nbsp;¼½ ÖÕcpDNASSR·Õ Ës2.3.1 相关系数 2维比较9 Ø4Õ Ë¥M1“ V AòÕ Ë“ Ð Ë ¥ z; Ö“ ¤¥“ Ë ²TÐM »“ Dice¥ Ë dÈÚ,M1“ rij¹-0.984 44;ÐJ¤¥ Ë ¥M1“ rij¹-0.951 74;ÐSM¤¥ Ë ¥M1“ rijK®,9r -0.923 36,t“ rij¥ &#39;´ûÚ¿0.9,&#39;ñûrdÈz¥)Y。2-wayManteltest²TV ü ˲T ?zQ: c¥cpSSR·Õ&#39; f , V AòM1“ I³ ˲TdÈM »。2.3.2 树状图聚类情况ʨ¥+scpSSRþNTCP9_©¥48ñ ¼½ ¡ ÖÕ“ Ö¥cpDNA ˹14ñ, Ï13ñ Ë ÿ, î_© Ë ¥92.86%。sY¨Nei&#39;s(1972)L. Ö“ 、DiceÄ ¥“ 、Jaccard“ SM Ä ¥“ ¤¥ ˲TÉ1 , V?C Ù ò“ ¥´ÐM,¨Nei&#39;s“ 、D“ 、J“ ¤M¥ ˲T,SM“ ¤¥ ˲TM »,Ç +) µ,vÁ|48z ¼½ ks¹ BF:»BF:v sl&#39;。»=F:e£&#39;、B×e£&#39;、ëÀ&#39;、B Ä&#39;、v½&#39; ¼、õ*â&#39;、õv £&#39;、 sl&#39;、õK&#39;、 A+&#39;、ϯK¥&#39;、 ´&#39;,ö1¹ ¼、 ÒÁ ¼½。» ØF:Ö×7 ¼&#39;、 2Ø*£&#39;、 Ø ÖÜ&#39;、lÖÜ&#39;、m öâ&#39;、¥&#39;、Ö &#39;、x&#39;、*x&#39;、 2ë é&#39;、 $ T&#39;、Ö×Õ&#39;、ª&#39;、 K Å ¦&#39;、%¥ Â=&#39;、½ Â=&#39;,ö1¹ÖÜ、x ¼ Ù ¼ Ë。» ¹F:FÜKl&#39;、 =U&#39;、Å&lt;¥&#39;、 ö&lt;¥&#39;、ÖÍ&lt;¥&#39;、x&lt;¥&#39;、x½äõ&lt;¥&#39;、&lt;¥&#39;、 släõ&lt;¥&#39;、 C Ê&lt;¥&#39;、Ö&lt;¥&#39;、5&lt;¥&#39;、Æ 3&lt;¥&#39;,ö1¹&lt;¥ ¼ Ë。»ôF: §&#39;、* sl&#39;、v&#39;、äõûî&#39;、l£É¶&#39;。» BF: 1&lt;¥&#39;。ÐAFLP ˲TM1 &nbsp;V?C;µM­),9µ­)。K üA¥­)¿cpSSRS:¥cpDNA¥L.M » üAÚ¿AFLPS:¥DNA¥L.M »,µ ÖÕ¥M »“ r1。iV üt ¼½ ¡ ÖÕ¥= s8yü M,7 ·¾+sþ 9Hq/,= s8yF¥M » Ú,9ü ª ÷ Ú,Mº­W¥µs9¸ ð+sþHq/?kY。î ðy V ? ¼½¥= s8yF ÖÕ £ Ü µ Ú¥÷,cpDNA¥L.? p。3 遗传相似系数分析与讨论ô &nbsp;48z ¼½ cpSSR ÿ9 Ø L.M »“ ,²T V©, k¥48ñ ¼½ ÖÕWL.M »“ ¥M¹0.400 1.000, Ü (L.M»“ ¹0.806。 Ï, ¼ ¼½ ÖÕWL.M »“ ¥M¹0.5331.000, Ü (L.M »“ ¹0.824; sl ¼½ ÖÕWL.M »“¥M¹0.5330.667, Ü (L.M »“ ¹0.600; ÒÁ ¼½ ÖÕWL.M »“ ¥M¹0.8001.000, Ü (L.M »“ ¹0.889;x ¼½ ÖÕWL.M »“ ¥M¹0.8671.000, Ü (L.M »“ ¹0.933;ÖÜ ¼½ÖÕWL.M »“ ¥M¹0.7331.000, Ü(L.M »“ ¹0.880;ë é ¼½ ÖÕWL.M »“ ¥M¹0.667 1.000, Ü (L.M»“ ¹0.773;&lt;¥ ¼ Ë ÖÕWL.M »“¥M¹0.7331.000, Ü (L.M »“ ¹0.926; ¼ Ù ¼ Ë ¼½ ÖÕWL.M »“ ¥M¹0.800 1.000, Ü (L.M »“ ¹0.907。 1&lt;¥&#39;、v sl&#39;¥= s8DNAÐ ð ¼½ ÖÕµv¥µs,V ü ñ ÖÕMsÄ *, ð S¥ ÖÕ。 ¼ ¼½ ÖÕ¥M »“ MKv,V üñ Ë¥ ¼½ ÖÕ ÷ *,ÉÄ å N É。ô &nbsp;48ñ ¼½ ¡ ÖÕ¥cpDNASSRs, V A ¼½ ¡ ÖÕW Ù iBç¥cpDNAMs, L.1“ ¯ Í, ª ü= s8DNA¥ &nbsp;o×ç,Q ¼½ ¡ ÖÕWy ÔN¥+Ä。®þNTCP9_©µ¥ ¼½ ¡ ÖÕcpDNAL.M »r 1.000, è  A+&#39;、õv £&#39;、v½ ¼&#39;ÐB Ä&#39;©, ñ Ì V ? LµL.M »¥ß, = s8DNA ¥µs¾þ/ ?z¥_©。 VnÇYVBÕ+sþ ÉcpSSRùî±þ ÖÕW¥ Tø1“ ¥。 ²TsV ü, ¼½ ¡ ÖÕW¥cpDNAµ Ú¥L.M »,9iBç¥L.µs9 &lt;1 g ö©: ¼½ ÖÕ&#39;÷= s8yFSSRùî53 sÄ。 ˲T&#39;Cµ¥s Ë“d,¯ iBç¥ f , ª ü ¼½ ¡ ÖÕ¥L.ß1 ¯,w© ÉÄ、 ¡âVñÏ, û Ú¥L.×ç¥ H,µBç¥L.Ms,®¿= s8DNA ×ç¥ TL., V ?£U ÖÕW÷* ùBt¥ Tø1“。 6,cpSSR¥ÖÕkY ? ï ÂAFLP, V4= s8y TL.,¹ùî ¼½ ÖÕ¥ Tø1“4 ¥± I。Á +YË 2ØÏ ö 3 ¼½ ö¥ B óÚý, 2Ø j&lt;vÐóÐý¿ ö¬ q,óq öÏS ¼½ùîÏ ° i öÚý、foö ©k §Úý“ ¼½“ Ö HóÍ¥ £ fúù¦û。 &nbsp;I Ó D 1 JURGEF.GeneticflexibilityofplantchloroplastJ.Nature,1999, 398:115-116. 2 WOLFEKH, LIWH,SHARPPM.Ratesofnucleotidesubstitu-tionvarygreatlyamongplantmitochondrial, chloroplast, andnu-clearDNAsJ.ProcNatlAcadSciUSA, 1987, 84(24):9 054-9 058. 3 CHENGYJ, GUOWW, DENGXX.cpSSR:Anewtooltoan-alyzechlorplastgenomeofcitrussomatichybridsJ.ActaBotani-caSinica, 2003, 45(8):906-909. 4 PLOMIONC, LEPROVOSTG, POTD, etal.Polencontamina-tioninamaritimepinepolycrossseedorchardandcertificationofimprovedseedsusingchloroplastmicrosatelitesJ.CanadianJournalofForestResearch, 2001, 31(10):1 815-1 816. 5 VIARDF, EL-KASSBYYA, RITLANDK.DiversityandgeneticstructureinpopulationsofPseudotsugamenziesi(Pinaceae)atchloroplastmicrosatelitelociJ.Genome, 2001, 44(3):336-344. 6 FERRISFCR,KINGRA,HEWITTGM,etal.ChloroplastDNArecognizesthreerefugialsourcesofEuropeanoaksandsuggestsin-dependenteasternandwesternimmigrationstoKnlandJ.Hered-ity, 1998, 80(5):584-593. 7 ISODAK, SHIRAISHIS, WATANABES, etal.Molecularevi-denceofnaturalhybridizationbetween, AbiesveitdhiandA.homolepis(Pinaceae)revealedbychlorplast, mitochondrialandnuclearDNAmarkersJ.MolEcol, 2000, 9(12):1 965-1 974. 8 JASONWL,MICHAELJH, SCOTTAJ, etal.CytogenomicanalysesrevealthestructuralplasticityofthechloroplastgenomeinhigherplantsJ.PlantCel, 2001, 13:245-254. 9 KATOS,YAMAGACHH, SHIMAOTOY, etal.Thechloroplastgenomesofazukibeananditscloserelatives:AdeletionmutationfoundinweedazukibeanJ.Hereditas, 2000, 132(1):43-48.10 ROSAA, FRANCOISL, MARIAT, etal.Chloroplastmicrosatel-litepolymorphismsinVitisspecies.J.Genome, 2002, 45(6):1 142-1 149.11 RICHARDGO,JEFFREYDP.ChloroplastDNAsystematics:AreviewofmethodsanddataanalysisJ.AmerJBot, 1994, 81:1 205-1 224.12 SCOTTNS, POSSINGHAMJV.ChangesinchloroplastDNAlev-elsduringgrowthofspinachleavesJ.JExpBot, 1983,34:1 756-1 767.13¢ç1, ÙTÝ,Û¡Ó. ¦% ÖÕSSRs¥ùîJ.óq±þÐùî, 2003, 21(5):444-448.14 ´¿,f ,ô,ñ¹ ¦.SSRs0S:¥ 7?/ ùîÉZJ. 2 j&lt;Ð, 2002, 15(4):106-109.15ñ W½, gZ,?,©.= s8DNA Ë ¥RFLP¥sÛÚB“dÐùîÏ¥¨J.±þs ËÐ, 2000, 38(2):97-110.16çu¬, &nbsp; 3, p. õ !k õ 8¥= s8±ÏsJ. &lt; SÐ, 2000, 36(3):46-51.17f=,â FÉ,Û ù ¥. °&#39; = ¾ 8¥= s8SSRsJ.L., 2004, 26(4):486-490.18 Ê ¨7. æ¨SSRkç £Õ0B¥ùîJ.Õ0,2003(1):23-25.19 ® ù, d b,· .:. &nbsp;2l 、ö7 3l # ôYl = s8DNAKÅ = M ½m ö¥ùîJ.L., 1995, 17(6):4-6.20· XÐ. 3þ ÐM.Ø: SÐñ, 1999.21ç .É.ÏS ¼½ ÖÕm½M.Ø:ÏS &lt;ñ, 1989.22¢î !.±þs0S:ð ØÐZEM.É ì:µ 2 SÐ/ ñ, 2003.(责任编辑 李 慧)</p>